CN111588801B

CN111588801B - Traditional Chinese medicine dispersible granule for preventing livestock and poultry epidemic diseases and preparation method thereof

Info

Publication number: CN111588801B
Application number: CN202010516974.0A
Authority: CN
Inventors: 杨海明
Original assignee: Yuanheng Biotechnology Co ltd
Current assignee: Yuanheng Biotechnology Co ltd
Priority date: 2020-06-09
Filing date: 2020-06-09
Publication date: 2021-10-01
Anticipated expiration: 2040-06-09
Also published as: CN111588801A

Abstract

The invention provides a traditional Chinese medicine dispersible granule for preventing livestock and poultry epidemic diseases and a preparation method thereof, belonging to the technical field of veterinary medicines. The traditional Chinese medicine dispersible granule mainly comprises 125 parts of 115-125 parts of rheum officinale, 38-42 parts of liquorice, 125 parts of 115-125 parts of scutellaria baicalensis, 208 parts of salvia miltiorrhiza, 38-42 parts of acanthopanax, 125 parts of 115-125 parts of trichosanthes root, 36-40 parts of sweet wormwood herb, 38-42 parts of schizonepeta, 125 parts of 115-125 parts of radix ophiopogonis, 52-58 parts of isatis root, 60-66 parts of bighead atractylodes rhizome and 125 parts of 115-125 parts of talcum. The invention applies the syndrome differentiation principle of the traditional Chinese medicine, has reasonable compatibility of medicines, fully exerts the function of each medicine, and coordinates all the medicines together to gain the effect of the medicine. The invention has the functions of regulating the organism immunity of livestock and poultry, enhancing the immune response capability of livestock and poultry and improving the capability of livestock and poultry for resisting epidemic diseases; the invention is a pure Chinese medicinal preparation, has no problem of antibiotic residue, reduces the influence on environmental pollution, and is safe and nontoxic; meanwhile, the preparation method disclosed by the invention is simple in preparation process, strong in product quality stability, long in storage time, controllable in quality standard, capable of realizing quantitative production and good in market popularization and application.

Description

Traditional Chinese medicine dispersible granule for preventing livestock and poultry epidemic diseases and preparation method thereof

Technical Field

The invention belongs to the technical field of veterinary drugs, and particularly relates to a traditional Chinese medicine dispersible granule for preventing livestock and poultry epidemic diseases and a preparation method thereof.

Background

In recent years, the development of livestock husbandry in China is leaps and bounds, the proportion of the livestock husbandry in the total agricultural value is improved year by year, and the livestock husbandry is gradually a supporting industry for guaranteeing food safety and promoting the income increase of farmers. The enlargement of the livestock and poultry breeding scale and the increase of epidemic diseases, and the production of some chemical treatment medicines such as antibiotics and hormones with rapid operation and definite curative effect is widely used and rapidly popularized, thereby playing a very important role in preventing and controlling animal diseases. However, the problems of the drug resistance of bacteria, the drug residue of animal products and the like are increasingly highlighted due to long-term, large-amount and unreasonable use of chemical drugs. Therefore, many people are aware of the defects and shortcomings of the chemical, and gradually turn the eyes to Chinese herbal medicines which are used as livestock and poultry health care products. The traditional Chinese medicine has the advantages of low toxicity, low residue, greenness, safety and the like, and a large number of researches report that the Chinese herbal medicine has obvious effects of resisting bacteria and viruses, regulating immunity, regulating nutrition and the like, so that the defects of chemical medicines are gradually applied to the prevention and treatment of livestock and poultry epidemic diseases at present.

Of the epidemic diseases of livestock and poultry, newcastle disease and swine fever are the most common and recurrent. Among them, newcastle disease is a contagious infection of birds by viruses, and is characterized by acute high incidence. The newcastle disease virus is caused by a virus belonging to the genus mumps of the family paramyxoviridae, and can invade the body of an avian through the respiratory tract, the digestive tract, the conjunctiva of the eyes, the wound and the like. The swine fever is one of 16 legal infectious diseases of A class marked by the world animal health organization, and has extremely strong infectivity and extremely high mortality. If large-scale swine fever occurs in the live pig breeding process, not only can the breeding economic benefit of farmers be directly reduced, but also serious problems of food safety, environmental pollution, animal epidemic disease prevention and control and the like can be caused. The swine fever belongs to the genus of pestivirus of the family flaviviridae, and has strong infectivity, a latent period of five to seven days generally and strong virus adaptability.

The species and serotype of the livestock and poultry virus are complex and are easy to mutate, and some viruses can cause an immune mechanism, which brings difficulty to vaccine research and application. The antiviral western medicines are few in types, large in toxic and side effects, easy to generate drug resistance and very limited in clinical curative effect. Most of the traditional Chinese medicines are natural plants and mineral substances, and most of the traditional Chinese medicines are substances which are beneficial and harmless to the organism through repeated tests and screening of human beings, and many of the traditional Chinese medicines are also varieties which can be used as food and medicine, although part of the traditional Chinese medicines have certain toxicity, the toxicity can be reduced through proper processing and compatibility, and the traditional Chinese medicines are nontoxic when being used in proper amount. The main components of the traditional Chinese medicine are biological macromolecules such as alkaloid, volatile oil, glycosides, organic acid, polysaccharide, tannin and the like, the macromolecular substances are combined in a compound form, complex metabolic reaction is realized in an organism, the survival and reproduction succession of pathogenic microorganisms is interfered or blocked from a plurality of links such as respiration, biochemical metabolism, energy conversion, genetic substances, reproduction and the like after entering the organism, and the pathogenic microorganisms are difficult to generate drug resistance to the pathogenic microorganisms, so that the pathogenic microorganisms are effectively inhibited and killed.

In the prior art, most of the traditional Chinese medicines for livestock and poultry have uncertain prescription sources, and the traditional Chinese medicine preparation is more in the form of oral liquid and powder. Wherein, the oral liquid is easy to be rancid and expand bottles, the powder is easy to be caked, and the prevention and treatment effect of the traditional Chinese medicine is greatly reduced. The bioavailability of the traditional Chinese medicine powder is generally low, the solubility of common traditional Chinese medicine granules and soaking decoction granules is relatively high in dependence on temperature, and the intensive winter cultivation is limited.

Disclosure of Invention

In order to solve the defects of the prior art, the invention aims to provide the traditional Chinese medicine dispersible granule for preventing the livestock and poultry epidemic disease and the preparation method thereof.

In order to realize the purpose of the invention, the technical scheme of the invention is as follows:

firstly, the invention provides a traditional Chinese medicine dispersion preparation for preventing livestock and poultry epidemic diseases, which comprises the following raw materials in parts by weight: rhubarb, liquorice, scutellaria, salvia, acanthopanax, trichosanthes root, sweet wormwood, schizonepeta, ophiopogon root, isatis root, bighead atractylodes rhizome and gypsum.

Further, as an optimization of the traditional Chinese medicine dispersion preparation for preventing the livestock and poultry epidemic diseases, the traditional Chinese medicine dispersion preparation comprises the following raw materials in parts by weight: 125 parts of rhubarb 115-containing material, 38-42 parts of liquorice, 125 parts of scutellaria 115-containing material, 208 parts of salvia 192-containing material, 38-42 parts of acanthopanax, 125 parts of trichosanthes 115-containing material, 36-40 parts of sweet wormwood, 38-42 parts of schizonepeta, 125 parts of dwarf lilyturf tuber 115-containing material, 52-58 parts of isatis root, 60-66 parts of bighead atractylodes rhizome and 125 parts of talc 115-containing material.

Further, as an optimization of the traditional Chinese medicine dispersion preparation for preventing the livestock and poultry epidemic diseases, the traditional Chinese medicine dispersion preparation comprises the following raw materials in parts by weight: 120 parts of rhubarb, 40 parts of liquorice, 115 parts of scutellaria, 204 parts of salvia miltiorrhiza, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root, 65 parts of bighead atractylodes rhizome and 115 parts of talcum.

Further, as the optimization of the traditional Chinese medicine dispersion preparation for preventing the livestock and poultry epidemic diseases, the traditional Chinese medicine dispersion preparation also comprises auxiliary materials, wherein the auxiliary materials comprise the following raw materials in parts by weight: 150 parts of polyvinylpyrrolidone 140-150 parts, 300 parts of lactose 290-60 parts, 60-70 parts of sodium bicarbonate, 70-80 parts of citric acid and 0.5-1 part of frankincense.

On the other hand, the invention provides a preparation method of a traditional Chinese medicine dispersion preparation for preventing livestock and poultry epidemic diseases, which is prepared according to the following steps:

(1) accurately weighing raw materials according to parts by weight of rhubarb, liquorice, scutellaria, salvia, acanthopanax, trichosanthes root, sweet wormwood, schizonepeta, ophiopogon root, isatis root, bighead atractylodes rhizome and talc respectively;

(2) putting the Chinese herbal medicine raw materials except the talc in the step (1) into a traditional Chinese medicine extractor, then adding 10-15 times of water in parts by weight of the raw materials, decocting for 30-40min, adding the talc, continuing to decoct for 30-40min, filtering and collecting filtrate, adding 8-10 times of water in parts by weight of the first total part into medicine dregs again, continuing to decoct for 60-70min, filtering and collecting filtrate, combining the filtrates collected twice, and performing spray drying to obtain a mixture A;

(3) accurately weighing polyvinylpyrrolidone, lactose, sodium bicarbonate, citric acid and porcine agalactia according to parts by weight, wherein the lactose, the sodium bicarbonate and the citric acid are respectively crushed and sieved by a 100-mesh and 200-mesh drug sieve; mixing the raw materials to obtain a mixture B;

(4) putting the mixture A and the mixture B into a mixing unit, and mixing for 45min to obtain a mixture C;

(5) and putting the mixture C into a dry tablet press for granules, tabletting, crushing, sieving by a No. 1 medicine sieve, and finishing granules to obtain the traditional Chinese medicine dispersible granules for preventing the livestock and poultry epidemic diseases.

The pharmacological action of the Chinese herbal medicine

Rhubarb: bitter taste and cold nature, entering stomach and liver meridians; has the effects of purging heat, cooling blood, removing toxicity, dispelling blood stasis, dredging channels and the like; has catharsis, cholagogue, hepatoprotective, antiulcer, antibacterial, and antiviral effects.

Licorice root: sweet in taste, neutral in nature, and entering heart, lung, spleen and stomach meridians; has the effects of invigorating spleen and replenishing qi, clearing away heat and toxic materials, eliminating phlegm and relieving cough, relieving spasm and pain, and harmonizing the medicines; it is often used for treating spleen and stomach weakness, asthenia, palpitation, short breath, cough, excessive phlegm, carbuncle, swelling, and sore, and can relieve drug toxicity and pungency.

Scutellaria baicalensis: bitter taste and cold nature, entering spleen and lung meridians; it contains baicalin, baicalein, wogonin I, wogonin II, norwogonin, volatile oil, and phenethyl alcohol glycoside, and has effects in clearing away heat and toxic materials, eliminating dampness, purging pathogenic fire, stopping bleeding, preventing miscarriage, lowering blood pressure, killing bacteria, killing virus, and killing fungi. The baicalin and baicalein contained in the scutellaria have good antioxidant function, and the scutellaria is used for treating ischemic stroke (cerebral infarction) and Parkinson's disease in human medicine and is used for treating foot-and-mouth disease and blister disease in veterinary medicine, clearing away lung heat and eliminating respiratory tract infection.

Red sage root: bitter taste, slightly cold nature, heart and liver meridians entered; has immunoregulation effect on cytokine, antibody, immune complex and immunocyte, and has effects of improving microcirculation, protecting liver, relieving inflammation, promoting blood circulation, removing blood stasis, tranquilizing mind, relieving pain, dilating blood vessel, and lowering blood pressure.

Acanthopanax root: sweet, slightly bitter and warm in nature; spleen, lung, heart and kidney meridians; has effects of invigorating qi, invigorating spleen, invigorating kidney, and tranquilizing mind, and can be used for treating spleen and lung qi deficiency, kidney deficiency, soreness of waist and knees, heart and spleen deficiency, and insomnia.

Trichosanthes root: sweet, slightly bitter and slightly cold in nature; entering lung and stomach meridians; has effects in clearing away heat, purging pathogenic fire, promoting salivation, quenching thirst, relieving swelling, and expelling pus; it is commonly used for treating feverish diseases with polydipsia, cough due to lung heat, diabetes due to internal heat, sores and ulcers, etc.

Sweet wormwood herb: bitter and pungent taste, cold in nature; has effects in clearing away deficiency heat, removing bone-steaming, relieving summer-heat, preventing malaria, and eliminating jaundice; it is commonly used for treating yin deficiency, fever due to yin deficiency, malaria, jaundice due to damp-heat, etc.; artemisia apiacea has antimalarial components. The discovery of artemisinin is a great contribution of China to mankind. Artemisia annua is used by veterinarians for the treatment of toxoplasmosis, eperythrozoonosis and viral diseases in animals.

Herba schizonepetae: pungent and bitter in flavor, slightly warm in nature, entering lung and liver meridians; has effects in relieving exterior syndrome, expelling pathogenic wind, promoting eruption, and stopping bleeding; the main active ingredients in the schizonepeta are volatile oil and flavonoid compounds, and the schizonepeta has obvious effects of sweating, relieving fever, resisting inflammation and easing pain; the effective components in herba Schizonepetae can inhibit the content of related inflammatory factors, shorten the cell growth cycle, and further exert antibacterial, antiviral, and antitumor effects.

Radix ophiopogonis: sweet and slightly bitter in taste, slightly cold in nature, entering heart, lung and stomach meridians; has the effects of nourishing yin, promoting the production of body fluid, moistening lung, relieving cough and the like; the chemical components of radix Ophiopogonis mainly include steroid saponin, homoisoflavonoid, polysaccharide, etc., and have pharmacological effects of lowering blood sugar, protecting cardiovascular system, enhancing immunity, resisting skin aging, resisting inflammation, resisting tumor, etc.

Radix isatidis: cold in nature, bitter in taste, entering heart and stomach meridians; has effects of clearing away heat and toxic materials, cooling blood, and relieving sore throat, and can be used for treating pestilence, toxin, carbuncle, macula, erysipelas, and wind-heat syndrome. It contains indigo, indirubin, beta-sitosterol, sinigrin, indioside, tryptanthrin, palmitic acid, sucrose, 1-thiocyanic acid-2-hydroxybut-3-ene and various amino acids, and the indioside is the main antiviral component. The radix Isatidis has strong antiviral, antibacterial, immunity enhancing, endotoxin resisting, anticancer and antipyretic effects.

White atractylodes rhizome: bitter and sweet in property, warm in nature, entering spleen and stomach meridians; has the effects of strengthening spleen, replenishing qi, eliminating dampness, promoting diuresis, arresting sweating, preventing miscarriage, etc.; it is often used for spleen deficiency, poor appetite, diarrhea, loose stool, threatened abortion, etc.; the main chemical components of the composition comprise volatile oil, polysaccharide, lactone and the like, and the composition has the effects of resisting tumors, resisting inflammation, regulating digestive systems and the like.

Talc: sweet in taste and cold in nature, and enters bladder, lung and stomach meridians; has effects in promoting urination, treating stranguria, clearing away summer-heat; it is commonly indicated for heat stranguria, urolithic stranguria, painful urination with heat, damp-heat diarrhea, etc.

Advantageous effects

The invention provides a traditional Chinese medicine dispersible granule for preventing livestock and poultry and a preparation method thereof. The invention is a pure Chinese medicinal preparation, mainly composed of eleven Chinese herbal medicines such as rhubarb, liquorice, scutellaria, salvia miltiorrhiza, acanthopanax, trichosanthes root and the like. The invention has the functions of regulating the organism immunity of livestock and poultry, enhancing the immune response capability of livestock and poultry and improving the capability of livestock and poultry for resisting epidemic diseases; the invention is a pure Chinese medicinal preparation, has no problem of antibiotic residue, reduces the influence on environmental pollution, and is safe and nontoxic; meanwhile, the preparation method has the advantages of simple preparation process, strong product quality stability, long storage time, controllable quality standard, capability of realizing quantitative production and good market popularization and application values.

Detailed Description

Example 1

A traditional Chinese medicine dispersible granule for preventing livestock and poultry epidemic diseases is prepared according to the following steps:

(1) the raw materials are accurately weighed respectively according to the parts by weight of 120 parts of rhubarb, 40 parts of liquorice, 115 parts of scutellaria, 204 parts of salvia, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root, 65 parts of atractylodes macrocephala and 115 parts of talc;

(2) putting all the Chinese herbal medicine raw materials except talc in the step (1) into a traditional Chinese medicine extractor, then adding 15 times of water in the sum of the weight parts of the raw materials, decocting for 40min, adding the talc, continuing to decoct for 40min, filtering and collecting filtrate, adding 10 times of water in the first total part into dregs of a decoction again, continuing to decoct for 70min, filtering and collecting filtrate, combining the filtrates collected twice, and performing spray drying to obtain a mixture A;

(3) respectively and accurately weighing 140 parts of polyvinylpyrrolidone, 290 parts of lactose, 60 parts of sodium bicarbonate, 70 parts of citric acid and 0.5 part of pig frankincense according to parts by weight, wherein the lactose, the sodium bicarbonate and the citric acid are respectively crushed and sieved by a 100-mesh medicine sieve; mixing the raw materials to obtain a mixture B;

The invention adopts the theory of traditional Chinese medicine, selects medicinal materials and makes the prescription of the medicinal materials, and because the experimental data is more, the prescription is adjusted and tested for many times, only a plurality of groups of prescriptions with higher curative effect are listed and compared with the prescription of the invention.

Formula 1: 120 parts of rhubarb, 115 parts of scutellaria, 204 parts of salvia, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 55 parts of isatis root, 65 parts of white atractylodes rhizome and 115 parts of talcum

And (2) formula: 120 parts of rhubarb, 40 parts of liquorice, 115 parts of scutellaria, 204 parts of salvia, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber and 115 parts of talc

And (3) formula: 120 parts of rhubarb, 40 parts of liquorice, 204 parts of salvia miltiorrhiza, 38 parts of acanthopanax, 118 parts of trichosanthes root, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root, 65 parts of largehead atractylodes rhizome and 115 parts of talcum

And (4) formula: 120 parts of rhubarb, 40 parts of liquorice, 115 parts of scutellaria, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root, 65 parts of white atractylodes rhizome and 115 parts of talc

And (5) formula: 40 parts of liquorice, 115 parts of scutellaria baicalensis, 204 parts of salvia miltiorrhiza, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood herb, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root, 65 parts of bighead atractylodes rhizome and 115 parts of talc

And (6) formula: 120 parts of rhubarb, 40 parts of liquorice, 115 parts of scutellaria, 204 parts of salvia, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root and 65 parts of bighead atractylodes rhizome

The formula of the invention is as follows: 120 parts of rhubarb, 40 parts of liquorice, 115 parts of scutellaria, 204 parts of salvia, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root, 65 parts of bighead atractylodes rhizome and 115 parts of talc

The test method comprises the following steps: 270 AA broilers of 1 day old were randomly divided into 9 groups, namely groups A-I, each group had 3 replicates, each replicate 10 broilers. Wherein group A is blank group, and is fed with basic daily ration and normal drinking water; the group B is a model group, and basic ration and normal drinking water are fed; the group C-I is a test group, and the basic ration and the formula 1-formula 6 obtained by the preparation method of the embodiment 1 with the mass fraction of 20 percent and the traditional Chinese medicine dispersible granule solution of the formula are respectively fed, the traditional Chinese medicine dispersible granule solution is fed for 10-30 days, and the normal drinking water is fed for the rest of time. The test period is 35 days, and the feeding conditions and the environment are kept the same. Performing first immunization on 7 days old Newcastle disease of group B-I, and performing eye drop nasal drip on IV attenuated vaccine of Newcastle disease by 2 times; the 21-day-old of the B-I groups were re-immunized with 4 times of water for the IV attenuated vaccine of Newcastle disease. And (5) feeding the seeds to 35 days old, stopping feeding and drinking water for 12 hours, and calculating the survival rate. The results are shown in Table 1.

TABLE 1 influence of Chinese medicinal dispersible granules of different formulas on survival rate of broiler chickens infected with Newcastle disease (only,%)

Categories	Group A	Group B	Group C	Group D	Group E	Group F	Group G	Group H	Group I
										Number of animals	30	30	30	30	30	30	30	30	30
Number of survivors	30	23	27	25	24	26	24	28	29
										Number of deaths	0	7	3	5	6	4	6	2	1
Survival rate	0	76.7^a	90.0^b	83.3^b	80.0	86.7^b	80.0	93.3^b	96.7^b

Note that comparison with group A^a P< 0.01, compared with group B^b P＜0. 05。

As can be seen from the results in Table 1, compared with the group B, the group C-I has a positive influence on the life rate of the newcastle disease infected broiler chickens, which indicates that the formulas 1 to 6 and the traditional Chinese medicine granules have a certain treatment effect on the newcastle disease infected broiler chickens. However, the formulas 1 to 6 lack individual traditional Chinese medicine raw materials in the traditional Chinese medicine granule, and the curative effect of the traditional Chinese medicine granule is not obvious. Compared with the group B, the survival rate of the group I, namely the traditional Chinese medicine granule group is obviously improved (P is less than 0.05), and is far higher than that of other formula preparation groups. The results show that the traditional Chinese medicine granules can obviously reduce the death rate of the broiler chicken infected with the Newcastle disease, improve the survival rate and have the life protection effect, eleven traditional Chinese medicines such as rhubarb, liquorice, scutellaria and the like are selected and used in the formula, the traditional Chinese medicine granules are reasonably combined by applying the traditional Chinese medicine dialectical treatment principle, the effects of all the medicines are fully exerted, the medicines are coordinated together to gain the medicine effect, and the components in the formula are not limited.

To further illustrate the stability, toxicity and effect on specific immune function of the dispersed particles of the present invention, animal experiments are provided below as an illustration. The following dispersion granules of the present invention were prepared in accordance with example 1.

1. Stability study

The test method comprises the following steps: the dispersible granules of the present invention were prepared according to example 1, packaged on the market, left for 24 months at a temperature of 25. + -. 2 ℃ and a relative humidity of 60. + -. 10%, and sampled and examined at the end of 0, 3, 6, 9, 12, 18 and 24 months, respectively, and the results are shown in Table 2.

TABLE 2 Long-term test results of the dispersible granule of the present invention

Time (moon)	Shape of	Solubility in water	Moisture (%)	Content (%)
					0	Brown yellow granules	Compliance with regulations	1.8	99.5
3	Brown yellow granules	Compliance with regulations	1.8	99.5
					6	Brown yellow granules	Compliance with regulations	1.9	99.5
9	Brown yellow granules	Compliance with regulations	1.9	99.4
					12	Brown yellow granules	Compliance with regulations	1.9	99.4
18	Brown yellow granules	Compliance with regulations	2.1	99.3
					24	Brown yellow granules	Compliance with regulations	2.1	99.3

The above results show that the dispersible granule of the invention is placed for 24 months under the conditions that the temperature is 25 +/-2 ℃ and the relative humidity is 60 +/-10%, and the sampling detection is carried out at the end of 0, 3, 6, 9, 12, 18 and 24 months respectively, and all detection indexes have no obvious change, which indicates that the dispersible granule of the invention has good stability and can be stored for a long time.

2. Acute toxicity test

2.1 half Lethal Dose (LD)₅₀) Measurement of

Taking 12 healthy Kunming mice with the weight of 16 +/-1 g, randomly dividing the mice into high, medium and low 3 dose groups, wherein each group comprises 4 mice, and the mice are raised in cages with half of male and female. Fasting was for 12 h before the test, without restriction of water. The traditional Chinese medicine dispersible granules are orally administrated, the administration dosage of the high, medium and low dosage groups is respectively 5000 mg/kg BW, 1000 mg/kg BW and 200 mg/kg BW, the mice are normally fed after the stomach irrigation, the observation is carried out for 7 days, and the clinical symptoms and death conditions of the mice in each group are carefully recorded. If death occurs in the high dose group, the median Lethal Dose (LD) is determined by the simplified kouynian method₅₀) (ii) a If the mice in the high-dose group do not die, indicating that the toxicity of the product is low, the acute toxicity is analyzed by Maximum Tolerated Dose (MTD).

2.2 Maximum Tolerated Dose (MTD) test

40 healthy Kunming mice with the weight of 16 +/-1 g are randomly divided into a high test group, a middle test group and a low test group and a normal saline control group, wherein each group comprises 10 mice, and the mice are bred in cages with half of males and females. Fasting was for 12 h before the test, without restriction of water. Because the maximum concentration of the aqueous solution of the traditional Chinese medicine dispersible granule and the maximum gastric perfusion volume of a mouse are limited, the maximum dosage is 60000mg/kg BW, the gastric perfusion dosages of a high test group, a middle test group and a low test group are respectively 60000mg/kg BW, 30000 mg/kg BW and 15000 mg/kg BW, the normal saline with the same volume is perfused into a normal saline control group, and the normal saline is normally fed after the gastric perfusion and observed for 7 days. The observation item indicators are: behavioral and behavioral changes, respiratory changes, skin and hair changes, intake and drinking water changes, mental states, etc. in mice. The death of each group of mice was recorded, and the Maximum Tolerated Dose (MTD) was taken as the highest dose at which no death occurred, and the fold maximum tolerated dose was calculated.

Calculating the formula:

maximum tolerance times = (maximum tolerance/average body weight of mouse) × (average body weight of mouse/dose of livestock and poultry per time)

2.3 measurement of organ index

MTD test mice after the test was completed all mice were sacrificed, necropsied, heart, kidney, liver and spleen were isolated, weight of each organ was weighed, and index of each organ was recorded and calculated.

Calculating the formula: organ index = organ weight/mouse body weight

2.4 data processing

Data were processed using SPSS 17.0 and expressed as mean ± standard deviation.

2.5 results and analysis

At LD₅₀In the measurement test, no death occurred in each group of mice 7 days after gavage, so the MTD test was performed. In the MTD test, the death condition of each group of mice after 7 days of gastric lavage is shown in table 3, and the index of each organ of the mice is shown in table 4.

TABLE 3 mortality in MTD assay in groups of mice

Group of	Number of animals (only)	Administration dose (mg/kg BW)	Death number (only)
				High dose group	10	60000	0
Middle dose group	10	30000	0
				Low dose group	10	15000	0
Physiological saline control group	10	—	0

TABLE 4 major organ index in MTD test mice

Group of	Index of heart	Liver index	Renal index	Spleen index
					High dose group	6.57±1.31	43.6±5.23	14.5±2.14	2.48±0.12
Middle dose group	7.12±2.06	42.8±4.62	14.7±3.08	3.14±0.67
					Low dose group	6.75±1.49	43.1±5.20	15.6±2.27	2.71±0.82
Physiological saline control group	6.44±1.17	43.5±4.51	15.2±2.51	2.83±0.54

The results in table 3 show that the mice in the test group and the control group do not die, the maximum tolerance of the dispersible granule of the invention to the mice is determined to be 60000mg/kg BW, and the maximum tolerance multiple is calculated to be 150 times according to the maximum tolerance multiple formula, which indicates that the dispersible granule of the invention has higher safety to livestock and poultry and is an actual nontoxic medicament within the normal dosage range.

The observation of the mouse shows that after the intragastric administration is carried out for 0.5h, the respiratory frequency of the mice in the test group is accelerated, the hair is erected, and the body is micro-trembled; the normal saline control group mice are accelerated in respiratory rate and are dull. After 1.5h, the mice in the test group are curled into a pile, the spirit is depressed, and the body tremor disappears; the saline control mice began to eat and slowed down. After 1d, the mice in the high-dose group begin to take food and drink water, the mental state is improved, and the feces are not shaped; the mice in the medium and low dose groups and the normal saline control group recover to be normal. After 2 days, the feces and other indexes of the high-dose mice are recovered to be normal. No death occurred in 7d groups of mice, and no abnormality was observed in the organs due to the sacrifice of the major organs. The results of the main organ indexes of the mice show that the main organ indexes of the mice in the high, medium and low dose groups of the dispersible granules have no obvious difference compared with the normal saline control group, and the dispersed granules do not cause the change of the organs of the mice. The results show that the traditional Chinese medicine dispersible granule preparation is safe and nontoxic, has no harm to the organism and can be used in cultivation without worry.

3 the influence of the traditional Chinese medicine dispersible granules on the specific immunologic function of livestock and poultry

3.1 materials

3.1.1 test animals: 180 AA broilers of 1 day old; 30 three-way weaned piglets of 30 days old.

3.1.2 test drugs: 20 percent of the traditional Chinese medicine dispersible granule solution; levamisole hydrochloride tablets.

3.2 test methods

3.2.1 grouping of broiler chickens and determination of immune organ index and Newcastle disease antibody titer

180 AA broilers of 1 day old are randomly divided into 3 groups, namely a group I is a blank control group (normal saline), a group II is a levamisole hydrochloride group (10 mg/kg), a group III is a traditional Chinese medicine dispersible granule group (20 mass percent aqueous solution), each group is repeated for 3 times, and each group is repeated for 10 times. In each group, the first immunization of the Newcastle disease is carried out at the age of 7 days, and 2 times of IV attenuated vaccine of the Newcastle disease is instilled into eyes and nose; the 21-day-old Newcastle disease is reimmunized, and the Newcastle disease IV attenuated vaccine is 4 times of drinking water. The administration of each group is carried out by drinking water at a dose of 5-30 days old, and normal drinking water is fed in the rest time, the test period is 42 days, and the feeding conditions and the environment are kept the same. When the chickens are raised to the age of 32 days and 42 days, 10 chickens are randomly selected from each group, blood is collected from each wing vein by 1mL, and the chickens are kept standing to collect serum for detecting the antibody titer of the Newcastle disease. The detection method is operated according to the national standard Newcastle disease antibody detection technology GBT 16550-. When the chickens are raised to 42 days old, all the chickens are killed by bleeding, bursa of fabricius, thymus and spleen are taken, fat is removed, and the chickens are weighed and recorded. And calculating the indexes of bursa of Fabricius, thymus and spleen.

Immune organ index (g/kg) = immune organ weight (mg)/animal body weight (g)

3.2.2 ternary weaned piglet grouping and hog cholera vaccine immunization effect

30 ternary weaned piglets of 30 days old are randomly divided into three groups, namely a blank control group (normal saline) in the group I, a levamisole hydrochloride group (10 mg/kg) in the group II, and 10 dispersible granules (20 mass percent aqueous solution) in the group III. The groups are fed with water according to the dose at the age of 32 days and are continuously fed for 15 days, and the groups are injected with the normal dose of the swine fever live vaccine for immunization at the age of 35 days. The piglets are respectively subjected to 5mL blood collection on the ear veins of each group of piglets at the ages of 36 th day, 40 th day, 45 th day, 50 th day and 55 th day after vaccination, standing is carried out, serum is collected, and the swine fever serum antibody level is detected by adopting an enzyme-labeled analyzer.

3.3 results and discussion

3.3.1 Effect of the dispersible granule of the invention on immune organs of broilers

The detection results of the broiler chicken immune organs in each group are shown in table 5. The bursa index, spleen index and thymus index of the group II and the group III are all obviously higher than that of the group I (A)P<0.05), and the indexes of the immune organs of the group III are slightly improved compared with the group II, and the difference is obvious (P<0.05）。

TABLE 5 Effect of the dispersible granules of the present invention on immune organs of broiler chickens

Group of	Bursal index	Spleen index	Index of thymus
				Group I	1.62±0.01^a	1.16±0.03^a	1.80±0.10^a
Group II	1.74±0.02^b	1.24±0.02^b	2.26±0.04^b
				Group III	1.86±0.01^c	1.36±0.02^c	2.34±0.07^c

Note: the same column marked with different letters shows significant difference (P<0.05）。

The central immune organs of the poultry comprise bone marrow, thymus and bursa of fabricius, and the central immune organs induce the generation, proliferation, differentiation and maturation of immunocompetent cells and regulate the development of peripheral immune organs and the overall immune response of animals. Peripheral immune organs including spleen, bone marrow and mucosa-associated lymphoid tissue are sites for lymphocyte aggregation and immune response, thymus is a site for immature T cells to develop into mature T cells, bursa of fabricius is a site for B cell differentiation and maturation, and spleen contains a large amount of T lymphocytes and B lymphocytes. The immune organ is a traditional Chinese medicine place for realizing immune defense and immune regulation of animals, so the size of the immune organ index is closely related to the health of the broiler chickens. The results in table 5 show that the dispersible granules of the traditional Chinese medicine have positive effects on immune organ indexes of broiler chickens, and can obviously improve the immune organ indexes of infected broiler chickens, so that the dispersible granules of the traditional Chinese medicine can well promote the development of immune organs of organisms and enhance the immune capacity of the immune organs, and have good material basis for resisting microbial infection in hospital.

3.3.2 Effect of the dispersible granule of the invention on the Newcastle disease antibody titer of broiler chickens

Compared with the determination results of the newcastle disease antibody titer of the 32-day-old broiler chickens, the antibody titers of the group II and the group III are obviously higher than those of the group I, and the difference is obviousP<0.05); the Newcastle disease antibody titer of the broiler chickens of 42 days old, the antibody titers of the group II and the group III are obviously higher than that of the group I, and the difference is extremely obvious (P<0.05), and the antibody titer of the group III is obviously higher than that of the group II, and the difference is obvious (P<0.05). The results are detailed in Table 6

TABLE 6 influence of the Chinese medicinal dispersible granules on the Newcastle disease antibody titer of broiler chickens

Categories	Group I	Group II	Group III
				32 day old chicken	4.91±0.61^a	5.53±0.72^b	5.79±0.32^b
42 day old chicken	5.20±0.41^a	6.52±0.51^b	7.13±0.26^b

Note: the same row is marked with different letters to show significant difference (P<0.05）。

As can be seen from Table 6, the antibody levels of Newcastle disease in 32 days old groups II and III, in which the serum antibody level of Newcastle disease was the highest, significantly increased with the vaccination of the vaccine, as compared with group I. The antibody level of the newcastle disease in the 42-day-old II group and III group is continuously increased, and the level of the newcastle disease antibody in serum in the III group is still the highest, which indicates that the traditional Chinese medicine dispersible granule can improve the titer of the anti-NDV hemagglutination inhibition antibody of the chicken, and the traditional Chinese medicine dispersible granule has longer duration for maintaining the high level of the newcastle disease antibody of the chicken, and is superior to the levamisole group.

3.3.3 Effect of the dispersible granule of the invention on the immunization effect of the piglet swine fever vaccine

As can be seen from Table 7, the antibody levels in groups II and III were higher than those in group I during the experiment, with a significant difference (P<0.05). Antibody levels in each group reached a maximum at 45 days of age, with group iii antibody levels being highest; at 50 days of age, there was a different decrease in antibody levels in each group, but the antibody levels in group III remained higher and differed significantly from those in groups I and II (seeP<0.05). The above results indicate that the dispersible granule of the present invention is effective in increasing the antibody level of swine fever and can maintain a long antibody peak-hour maintenance time.

TABLE 7 Effect of the present invention on the immunization effect of the piglet Swine fever vaccine

Age of day	Group I	Group II	Group III
				32 days	0.35± 0.05^a	0.46±0.02^b	0.46±0.03^b
For 40 days	0.42± 0.04^a	0.52±0.05^b	0.59±0.06^b
				Day 45	0.49±0.07^a	0.65±0.03^b	0.72±0.04^c
50 days	0.38±0.01^a	0.58±0.02^b	0.64±0.01^c
				Day 55	0.34±0.02^a	0.55±0.06^b	0.59±0.06^b

Example 2

This embodiment is different from embodiment 1 in that

Raw materials (by weight portion): 115 parts of rhubarb, 38 parts of liquorice, 115 parts of scutellaria, 192 parts of salvia miltiorrhiza, 38 parts of acanthopanax, 115 parts of trichosanthes root, 36 parts of sweet wormwood, 38 parts of schizonepeta, 115 parts of dwarf lilyturf tuber, 52 parts of isatis root, 60 parts of bighead atractylodes rhizome, 115 parts of talc, 140 parts of polyvinylpyrrolidone, 290 parts of lactose, 60 parts of sodium bicarbonate, 70 parts of citric acid and 0.5 part of pig frankincense.

Example 3

This embodiment is different from embodiment 1 in that

Raw materials (by weight portion): 125 parts of rhubarb, 42 parts of liquorice, 125 parts of scutellaria, 208 parts of salvia miltiorrhiza, 42 parts of acanthopanax, 125 parts of trichosanthes root, 40 parts of sweet wormwood, 42 parts of schizonepeta, 125 parts of dwarf lilyturf tuber, 58 parts of isatis root, 66 parts of bighead atractylodes rhizome, 125 parts of talc, 150 parts of polyvinylpyrrolidone, 300 parts of lactose, 70 parts of sodium bicarbonate, 80 parts of citric acid and 1 part of pig frankincense.

Example 4

This embodiment is different from embodiment 1 in that

Raw materials (by weight portion): 120 parts of rhubarb, 40 parts of liquorice, 120 parts of scutellaria, 200 parts of salvia miltiorrhiza, 40 parts of acanthopanax, 120 parts of trichosanthes root, 38 parts of sweet wormwood, 40 parts of schizonepeta, 120 parts of dwarf lilyturf tuber, 56 parts of isatis root, 64 parts of bighead atractylodes rhizome, 120 parts of talc, 145 parts of polyvinylpyrrolidone, 295 parts of lactose, 65 parts of sodium bicarbonate, 75 parts of citric acid and 0.8 part of pig milk fragrance.

Example 5

This embodiment is different from embodiment 1 in that

Raw materials (by weight portion): 118 parts of rhubarb, 42 parts of liquorice, 122 parts of scutellaria baicalensis, 196 parts of salvia miltiorrhiza, 38 parts of acanthopanax, 115 parts of trichosanthes root, 40 parts of sweet wormwood, 42 parts of schizonepeta, 124 parts of dwarf lilyturf tuber, 55 parts of isatis root, 62 parts of bighead atractylodes rhizome, 116 parts of talc, 142 parts of polyvinylpyrrolidone, 296 parts of lactose, 68 parts of sodium bicarbonate, 72 parts of citric acid and 0.6 part of pig frankincense.

Example 6

This embodiment is different from embodiment 1 in that

Raw materials (by weight portion): 122 parts of rhubarb, 40 parts of liquorice, 116 parts of scutellaria baicalensis, 202 parts of salvia miltiorrhiza, 38 parts of acanthopanax, 116 parts of trichosanthes root, 40 parts of sweet wormwood, 39 parts of schizonepeta, 118 parts of dwarf lilyturf tuber, 52 parts of isatis root, 66 parts of bighead atractylodes rhizome, 115 parts of talc, 148 parts of polyvinylpyrrolidone, 298 parts of lactose, 66 parts of sodium bicarbonate, 76 parts of citric acid and 1 part of pig frankincense.

Example 7

This embodiment is different from embodiment 1 in that

Raw materials (by weight portion): 116 parts of rhubarb, 40 parts of liquorice, 124 parts of scutellaria baicalensis, 206 parts of salvia miltiorrhiza, 42 parts of acanthopanax, 122 parts of trichosanthes root, 38 parts of sweet wormwood, 42 parts of schizonepeta, 124 parts of dwarf lilyturf tuber, 54 parts of isatis root, 64 parts of bighead atractylodes rhizome, 116 parts of talc, 148 parts of polyvinylpyrrolidone, 292 parts of lactose, 62 parts of sodium bicarbonate, 78 parts of citric acid and 0.9 part of pig milk fragrance.

The above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. The traditional Chinese medicine dispersion preparation for preventing the livestock and poultry epidemic diseases is characterized by being prepared from the following raw materials in parts by weight: 125 parts of rhubarb 115-containing material, 38-42 parts of liquorice, 125 parts of scutellaria 115-containing material, 208 parts of salvia 192-containing material, 38-42 parts of acanthopanax, 125 parts of trichosanthes 115-containing material, 36-40 parts of sweet wormwood, 38-42 parts of schizonepeta, 125 parts of dwarf lilyturf tuber 115-containing material, 52-58 parts of isatis root, 60-66 parts of bighead atractylodes rhizome and 125 parts of talc 115-containing material; the traditional Chinese medicine dispersion preparation also comprises the following auxiliary materials in parts by weight: 150 parts of polyvinylpyrrolidone 140-150 parts, 300 parts of lactose 290-60 parts, 60-70 parts of sodium bicarbonate, 70-80 parts of citric acid and 0.5-1 part of frankincense.

2. The traditional Chinese medicine dispersion preparation for preventing the livestock and poultry epidemic diseases according to claim 1, which is characterized by being prepared from the following raw materials in parts by weight: 120 parts of rhubarb, 40 parts of liquorice, 115 parts of scutellaria, 204 parts of salvia miltiorrhiza, 38 parts of acanthopanax, 118 parts of trichosanthes root, 36 parts of sweet wormwood, 42 parts of schizonepeta, 122 parts of dwarf lilyturf tuber, 55 parts of isatis root, 65 parts of bighead atractylodes rhizome and 115 parts of talcum.

3. The preparation method of the traditional Chinese medicine dispersion preparation for preventing the livestock and poultry epidemic disease as claimed in any one of claims 1 to 2, characterized in that the preparation method comprises the following steps:

(1) accurately weighing the following components in parts by weight: radix et rhizoma Rhei, Glycyrrhrizae radix, Scutellariae radix, Saviae Miltiorrhizae radix, radix Acanthopanacis Senticosi, Trichosanthis radix, herba Artemisiae Annuae, herba Schizonepetae, radix Ophiopogonis, radix Isatidis, Atractylodis rhizoma, and pulvis Talci;

(2) putting all the Chinese herbal medicine raw materials except talc in the step (1) into a traditional Chinese medicine extractor, then adding 10-15 times of water of the sum of the weight parts of the raw materials, decocting for 30-40min, adding the talc, continuing to decoct for 30-40min, filtering and collecting filtrate, adding 8-10 times of water of the sum of the weight parts of the raw materials into dregs of a decoction again, continuing to decoct for 60-70min, filtering and collecting filtrate, combining the filtrates collected twice, and performing spray drying to obtain a mixture A;

(3) accurately weighing the following components in parts by weight: polyvinylpyrrolidone, lactose, sodium bicarbonate, citric acid and pig frankincense, wherein the lactose, the sodium bicarbonate and the citric acid are respectively crushed and sieved by a 100-mesh and 200-mesh drug sieve; mixing the auxiliary materials to obtain a mixture B;